同步荧光光谱法结合分子对接研究甲氧苄啶与牛血清白蛋白的相互作用  

Study on the Interaction of Trimethoprim(TMP)with Bovine Serum Albumin(BSA)by Combining Synchronous Fluorescence Spectrometry with Molecular Docking

作　　者：杨莹 李佳伟 王书琴 杜萍 童文华 YANG Ying;LI Jia-wei;WANG Shu-qin;DU Ping;TONG Wen-hua(School of Biological Engineering,Sichuan University of Science and Engineering,Yibin 644000,China;Wuliangye Yibin Co.,Ltd.,Yibin 644007,China;Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,Qingdao 266000,China)

机构地区：[1]四川轻化工大学生物工程学院,中国宜宾644000 [2]四川宜宾五粮液集团有限公司,中国宜宾644007 [3]中国科学院青岛生物能源与过程研究所,中国青岛266101

出　　处：《湖南师范大学自然科学学报》2022年第3期103-108,116,共7页Journal of Natural Science of Hunan Normal University

基　　金：四川轻化工大学人才引进项目(2019RC31);五粮液集团有限公司博士后基金资助项目(294074);四川轻化工大学大学生创新创业训练计划项目(cx2021143,cx2021135)。

摘　　要：利用荧光光谱法和紫外分光光度法在模拟生理条件下研究牛血清白蛋白(BSA)和甲氧苄啶(TMP)的相互作用,应用Stern-Volmer方程、Lineweaver-Burk双倒数方程及Förster非辐射能量转移理论,计算获得3个不同温度下的猝灭常数(K_(SV))、结合常数(K_(q))、结合距离(r)及热力学常数(ΔH,ΔG,ΔS)等参数。结果表明:3个不同温度下,BSA和TMP均能发生反应生成络合物,属于静态猝灭;结合距离小于7 nm;同步荧光光谱显示TMP对BSA的构象产生影响,主要是对其中酪氨酸残基的影响更为显著,紫外吸收光谱中BSA的最大吸收峰向较短波长偏移,表明BSA与TMP分子之间形成了基态配合物;圆二色谱显示TMP主要结合在BSA的主要多肽链的氨基酸残基上;分子对接结果表明TMP和BSA之间结合的主要作用力是疏水作用力和π-π作用。The interaction between trimethoprim(TMP)and bovine serum albumin(BSA)was studied by fluorescence spectroscopy and ultraviolet absorption spectroscopy under simulated human physiological conditions.Based on the Stern-Volmer equation,Lineweaver-Burk double reciprocal equation and Forster non-radiative energy transfer theory,the quenching constants(K_(SV)),bimolecular quenching constants(K_(q)),binding distance(r)and thermodynamic constants(ΔH,ΔG,ΔS)were determined at three different temperatures.The results show that BSA can react with TMP to form complex at three different temperatures,which belongs to static quenching.And the binding distance(r)is less than 7 nm.The synchronous fluorescence spectrum showed that TMP had an effect on the conformation of BSA,especially on the tyrosine residue.The maximum absorption peak of BSA in the ultraviolet absorption spectrum shifted to the shorter wavelength,indicating that the ground state complex was formed between BSA and TMP.Circular dichroism showed that TMP was mainly bound to the amino acid residues of the main polypeptide chain of BSA.Molecular docking results reveal that the main binding forces between TMP and BSA molecules are hydrophobic interaction andπ-πinteraction.

关 键 词：甲氧苄啶 牛血清白蛋白 同步荧光光谱 圆二色谱 分子对接 

分 类 号：O657.3[理学—分析化学] TQ460.1[理学—化学]